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目的研究母体缺氧对新西兰胎兔颈动脉生长因子、细胞外基质表达及血管形态学的影响。方法同期怀孕的新西兰母兔[足月(30±1)d]随机分为对照组(21%O_2,n=5)、缺氧组(12%O_2,n=5),缺氧组于妊娠第10天开始采用缺氧箱进行缺氧暴露,每天2次,上、下午各4h,至妊娠第29天结束。妊娠第29天,母兔麻醉,剖宫取出胎兔,分离胎兔颈动脉,液氮中速冻后转移至-70℃保存备检,另保存胎兔颈动脉用于免疫组织化学检查和Masson染色。用免疫印迹检测缺氧诱导因子1α(HIF-1α)、转化生长因子β_1(TGF-β_1)、Smad3和结缔组织生长因子(CTGF)蛋白表达;实时聚合酶链反应(Real-time PCR)检测HIF-1α、TGF-β_1和CTGF mRNA表达水平;免疫组织化学技术检测颈动脉TGF-β_1、CTGF、Ⅰ型胶原蛋白(CollⅠ)、Ⅲ型胶原蛋白(CollⅢ)表达;Masson染色检测颈动脉胶原纤维含量及内中膜厚度。结果与对照组比较,母体缺氧组胎兔颈动脉HIF-1α(0.26±0.05比0.07±0.04)、TGF-β_1(0.68±0.08比0.51±0.11)、Smad3(0.47±0.04比0.26±0.02)和CTGF(0.81±0.09比0.53±0.12)蛋白表达增多,HIF-1α(1.62±0.37比1.00±0.18)、TGF-β_1(1.90±0.35比1.00±0.12)和CTGF(1.82±0.42比1.00±0.21)mRNA水平升高,及CollⅠ(0.35±0.02比0.27±0.02)、CollⅢ(0.26±0.02比0.22±0.03)表达增多(P<0.05或P<0.01);而Masson染色结果提示:胶原纤维含量、内膜和中膜厚度在两组之间的差异无统计学意义(均P>0.05)。结论母体缺氧上调胎兔颈动脉TGF-β_1、CTGF、CollⅠ、CollⅢ的表达。
Objective To investigate the effects of maternal hypoxia on carotid artery growth factor, extracellular matrix expression and vascular morphology in New Zealand rabbits. Methods Pregnant New Zealand females (term, 30 ± 1 d) were randomly divided into control group (21% O_2, n = 5) and hypoxia group (12% On the 10th day, the hypoxia tank was used for hypoxia exposure twice a day for 4 hours in the morning and afternoon, and ended in the 29th day of pregnancy. On the 29th day of gestation, the femur was anesthetized and the fetus was removed from the cesarean section. The carotid arteries of fetus were isolated, frozen in liquid nitrogen and transferred to -70 ℃ for storage. The fetal carotid artery was also preserved for immunohistochemistry and masson staining . The protein expressions of HIF-1α, TGF-β_1, Smad3 and CTGF were detected by Western blotting. Real-time PCR was used to detect the expression of HIF-1α, The expressions of TGF-β 1, CTGF, Coll Ⅰ and Collagen III in carotid arteries were detected by immunohistochemistry. Masson staining was used to detect the content of collagen fibers in carotid artery And intima-media thickness. Results Compared with the control group, the HIF-1α (0.26 ± 0.05 vs 0.07 ± 0.04), TGF-β1 (0.68 ± 0.08 vs 0.51 ± 0.11), Smad3 (0.47 ± 0.04 vs 0.26 ± 0.02) (1.62 ± 0.37 vs 1.00 ± 0.18), TGF-β_1 (1.90 ± 0.35 vs 1.00 ± 0.12), and CTGF (1.82 ± 0.42 vs 1.00 ± 0.21), and CTGF (0.81 ± 0.09 vs 0.53 ± 0.12) ) mRNA levels increased with Coll Ⅰ (0.35 ± 0.02 vs 0.27 ± 0.02) and Coll Ⅲ (0.26 ± 0.02 vs 0.22 ± 0.03) (P <0.05 or P <0.01). The results of Masson staining suggested that collagen fibers content, There was no significant difference in intima and media thickness between the two groups (all P> 0.05). Conclusion The maternal anoxia up-regulates the expression of TGF-β 1, CTGF, Coll Ⅰ and Coll Ⅲ in the carotid artery of the fetus.